Engineered hot-start Taq DNA polymerase variants and their fully premixed kits

CN122235110APending Publication Date: 2026-06-19BEIJING BAILIGE BIOTECHNOLOGY CO LTD +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BEIJING BAILIGE BIOTECHNOLOGY CO LTD
Filing Date
2026-05-22
Publication Date
2026-06-19

AI Technical Summary

Technical Problem

In existing fully premixed PCR systems, Taq DNA polymerase is prone to non-specific reactions under low temperature conditions and has insufficient compatibility with temperature-sensitive inhibitors, leading to non-specific degradation of the probe and affecting detection sensitivity and stability.

Method used

By inserting a heat-labile inteptide into Taq DNA polymerase using genetic engineering techniques, a conformational change is induced, which binds to the thermophilic endonuclease 1 domain, achieving a hot-start effect, retaining 5'→3' exonuclease activity and enhancing substrate selectivity.

Benefits of technology

This technology enables hot-start PCR without the need for temperature-sensitive inhibitors, improving the specificity and stability of PCR, enhancing enzyme activity and detection sensitivity, and solving the problem of long-term storage of fully premixed systems.

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Abstract

This invention discloses an engineered hot-start Taq DNA polymerase variant and its fully premixed kit. The engineered hot-start Taq DNA polymerase variant of this invention is obtained by replacing the amino acid sequence of the 5'–3' exonuclease domain of wild-type Taq DNA polymerase with SEQ ID NO. 3 and inserting SEQ ID NO. 2 between amino acids 505 and 506 of wild-type Taq DNA polymerase. This invention's Taq DNA polymerase variant uses genetic engineering techniques to insert a heat-labile intron domain into its thumb domain, inducing a conformational change in this region, thereby inhibiting DNA binding. This invention's Taq DNA polymerase variant also incorporates the endonuclease 1 domain of *Thermotrophic thermococcus* through genetic engineering, successfully conferring substrate selectivity.
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